LOCUS       sequence-86315-e        6478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EF1a-driven expression of VCre in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6478)
  AUTHORS   Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone
            J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H,
            Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, 
            Deisseroth K
  TITLE     Targeting cells with single vectors using multiple-feature Boolean 
            logic.
  JOURNAL   Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 
            2014 Jun 8.
  PUBMED    24908100
REFERENCE   2  (bases 1 to 6478)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.2996"; journalName: "Nat Methods"; date: "2014-07"; 
            volume: "11"; issue: "7"; pages: "763-72"
COMMENT     Sequence Label: pAAV-EF1a-vCre
FEATURES             Location/Qualifiers
     source          1..6478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    2605..3193
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2658..2678)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3076..3087)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3225..3701
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3424..3443)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3741..3881
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3741..3870
                     /label=AAV2 ITR
     rep_origin      3956..4411
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4043..4062)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4253..4274
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4428..4447)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4547..4569
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4607..4625)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4693..4797
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4798..4866,4867..5658)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5016..5035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5829..6417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6318..6337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cgccaccatg
     1441 attgagaacc agctgtcact gctgggagat ttttcaggag tccgacccga tgatgtcaag
     1501 accgccattc aggccgcaca gaagaagggc atcaacgtgg cagagaatga acagttcaaa
     1561 gccgcttttg agcacctgct gaacgagttc aagaagcggg aggaacggta ctctccaaat
     1621 actctgcgga gactggagag tgcttggacc tgctttgtgg attggtgtct ggctaaccac
     1681 aggcatagtc tgccagcaac acccgacact gtcgaggcat tctttatcga aagggccgag
     1741 gaactgcacc gcaataccct gagcgtgtac cgctgggcca tttccagagt gcatagggtc
     1801 gctggatgcc ccgacccttg tctggatatc tatgtggagg atagactgaa ggctattgca
     1861 cggaagaaag tgagagaggg cgaagcagtc aaacaggcca gccccttcaa cgaacagcac
     1921 ctgctgaagc tgacatcact gtggtacaga agcgacaaac tgctgctgag gcgaaacctg
     1981 gccctgctgg ccgtggctta tgagtccatg ctgcgagctt ctgaactggc aaatatccgc
     2041 gtctccgaca tggagctggc cggcgatggg accgctattc tgacaatccc tattactaag
     2101 accaaccatt ctggagagcc agacacttgc atcctgagtc aggatgtggt ctcactgctg
     2161 atggactaca ccgaagccgg caagctggac atgagctccg atgggttcct gtttgtggga
     2221 gtcagcaaac acaatacatg tatcaagccc aagaaagata aacagactgg cgaggtgctg
     2281 cataagccta ttaccacaaa aaccgtggag ggggtcttct attcagcctg ggaaacactg
     2341 gatctgggac ggcagggcgt gaagcctttt acagcacata gcgccagagt cggagcagca
     2401 caggacctgc tgaagaaagg atacaatact ctgcagatcc agcagtccgg caggtggtct
     2461 agtggggcta tggtggcacg ctatggacga gccattctgg ctcgagacgg agcaatggca
     2521 catagcaggg tcaagactcg gtcagcacca atgcagtggg gaaaggacga aaaggactaa
     2581 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2641 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2701 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2761 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2821 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2881 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2941 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3001 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     3061 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3121 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3181 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg
     3241 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     3301 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     3361 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     3421 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     3481 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     3541 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     3601 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     3661 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca
     3721 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3781 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3841 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3901 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3961 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4021 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4081 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4141 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4201 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4261 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     4321 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4381 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4441 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4501 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4561 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4621 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4681 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4741 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4801 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4861 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4921 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4981 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5041 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5101 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5161 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5221 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5281 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5341 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5401 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5461 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5521 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5581 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5641 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5701 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5761 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5821 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5881 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5941 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6001 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6061 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6121 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6181 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6241 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6301 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6361 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6421 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
